Device for whole blood separation

ABSTRACT

Separation of the cellular components of whole blood, or other biological fluid, from plasma or serum can be achieved for assay analysis. A device for facilitating separation can include, for example, a capillary tube that accurately draws target blood volume, a pad that chemically interacts with red-blood cells, such that the red blood cells become chemically and/or physically trapped within pad material, a mechanism for plasma recovery from the pad upon diffusion or active mixing, and a dropper tip that facilitates dispensing the mixture onto a test device. The treatment of the cellular components can be performed prior to contact with a buffer solution, so release of the cellular components into the buffer solution is reduced or prevented. Additional filtration can be provided to filter any remaining cellular components in the mixture.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of U.S. Provisional Application No.62/423,062, filed Nov. 16, 2016, and U.S. Provisional Application No.62/464,331, filed Feb. 27, 2017, the entirety of each of which isincorporated herein by reference.

TECHNICAL FIELD

The present disclosure generally relates to separation of cellularcomponents from a biological sample, such as separation of whole bloodinto blood cells and plasma.

BACKGROUND

Various techniques can be employed to analyze body fluids for thepresence or absence of a particular soluble constituent. For example,tests are available to detect glucose, uric acid or protein in urine, orto detect glucose, triglycerides, potassium ion or cholesterol in blood.Devices and methods to process body fluids prior to testing are used,for example, to separate the fluid sample into separate portions orfractions. For example, cellular components of whole blood can beseparated from the plasma or serum for use in an assay to test for aparticular soluble constituent. The resulting plasma or serum can beexamined for a response to provide a prompt and accurate qualitative orquantitative assay for one or more soluble constituents of the plasma orserum.

BRIEF SUMMARY

The subject technology is illustrated, for example, according to variousaspects described below.

According to some embodiments of the present disclosure, a kit caninclude: a container including a reservoir for receiving a solution; asample device including: an inflow chamber for receiving a liquidsample; a mixing chamber; a pad between the inflow chamber and themixing chamber, the pad optionally including a processing reagent suchas a red blood cell agglomerating substance; an outflow port adjacent tothe mixing chamber; and an interface for sealably connecting the sampledevice to the container such that the reservoir is in fluidcommunication with the mixing chamber.

The inflow chamber can include a capillary tube. The sample devicefurther can include a filter between the mixing chamber and the outflowport. The kit can further include a solution within the reservoir.

According to some embodiments of the present disclosure, a method caninclude: with an inflow chamber of a sample device, receiving a liquidsample; facilitating separation of a first portion of the liquid samplefrom a second portion of the liquid sample by retaining the secondportion at a pad, the pad optionally including a processing reagent suchas red blood cell agglomerating substance when the liquid sample is, forexample, blood; facilitating flow of the first portion of the liquidsample through the pad and to a mixing chamber; sealably connecting thesample device to a container including a reservoir containing a buffersolution; mixing the first portion of the liquid sample with the buffersolution to create a mixture; and dispensing at least some of themixture from the mixing chamber and through an outflow port of thesample device.

The dispensing can include deforming at least a portion of the sampledevice or the container to increase a pressure within the mixingchamber. The dispensing can include facilitating flow through a filterbetween the mixing chamber and the outflow port. The dispensing can alsocomprise applying a directional force to the sample device or thecontainer to effect release of at least some of the mixture.

According to some embodiments of the present disclosure, a kit caninclude: a sample device including: a pad for receiving a liquid sample,the pad optionally including a processing reagent, such as a red bloodcell agglomerating substance; and a chamber having an outflow port; anda container including: a reservoir containing or configured to contain asolution; and an interface for sealably connecting the sample device tothe container such that the reservoir is in fluid communication with thechamber.

The kit can further include a collection device for collecting theliquid sample, such as a blood sample or a urine sample or a sputumsample from a patient. The collection device can include a capillarytube and a bulb in fluid communication with the capillary tube.

According to some embodiments of the present disclosure, a method caninclude: with a pad of a sample device, receiving a liquid sample, thepad optionally including a processing reagent such as a red blood cellagglomerating substance; sealably connecting the sample device to acontainer including a reservoir containing or configured to contain asolution; facilitating separation of a first portion of the liquidsample from a second portion of the liquid sample by retaining thesecond portion at the pad; mixing the first portion of the liquid samplewith the solution to create a mixture in a chamber of the sample device;and dispensing at least some of the mixture from the chamber and throughan outflow port of the sample device.

The dispensing can include deforming at least a portion of the sampledevice or the container to increase a pressure within the chamber. Thedispensing can include facilitating flow through a filter between thechamber and the outflow port. The dispensing can also comprise applyinga directional force to the sample device or the container to effectrelease of at least some of the mixture. The method can further includecollecting the liquid sample with a capillary tube of a collectiondevice for collecting the liquid sample, for example a blood sample froma patient, and receiving the liquid sample can include dispensing thesample from the capillary tube by deforming at least a portion of a bulbin fluid communication with the capillary tube.

According to some embodiments of the present disclosure, a kit caninclude: a container including: a reservoir containing or configured tocontain a solution; a mixing chamber; a barrier separating the reservoirfrom the mixing chamber; a sample device including: an inflow chamberfor receiving a liquid sample; a channel; a pad between the inflowchamber and the channel, the pad optionally including a processingreagent such as a red blood cell agglomerating substance; a dispenserincluding: an outflow port; an interface for sealably connecting thedispenser to the container such that the mixing chamber is in fluidcommunication with the outflow port, wherein the dispenser is configuredto urge the sample device through the barrier when the dispenser isconnected to the container.

The inflow chamber can include a capillary tube. The dispenser furthercan include a filter that is between the mixing chamber and the outflowport when the dispenser is connected to the container. The container caninclude at least one protrusion to secure at least a portion of thesample device within the mixing chamber.

According to some embodiments of the present disclosure, a method caninclude: with an inflow chamber of a sample device, receiving a liquidsample while at least a portion of the sample device is within acontainer; facilitating separation of a first portion of the liquidsample from a second portion of the liquid sample by retaining thesecond portion at a pad, the pad optionally including a processingreagent such as a red blood cell agglomerating substance; with adispenser, urging the sample device through a barrier separating areservoir of the container from a mixing chamber of the container;sealably connecting the dispenser to the container; mixing the firstportion of the liquid sample with a solution in the reservoir to createa mixture; and dispensing at least some of the mixture from the mixingchamber and through an outflow port of the sample device.

The dispensing can include deforming at least a portion of the containerto increase a pressure within the mixing chamber. The dispensing caninclude facilitating flow through a filter between the mixing chamberand the outflow port. The dispensing can also comprise applying adirectional force to the sample device or the container to effectrelease of at least some of the mixture.

In addition to the exemplary aspects and embodiments described above,further aspects and embodiments will become apparent by reference to thedrawings and by study of the following descriptions.

Additional embodiments of the present methods and compositions, and thelike, will be apparent from the following description, drawings,examples, and claims. As can be appreciated from the foregoing andfollowing description, each and every feature described herein, and eachand every combination of two or more of such features, is includedwithin the scope of the present disclosure provided that the featuresincluded in such a combination are not mutually inconsistent. Inaddition, any feature or combination of features may be specificallyexcluded from any embodiment of the present invention. Additionalaspects and advantages of the present invention are set forth in thefollowing description and claims, particularly when considered inconjunction with the accompanying examples and drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 illustrates a perspective view of a sample device, according tosome embodiments of the present disclosure.

FIG. 2 illustrates a sectional view of the sample device of FIG. 1,according to some embodiments of the present disclosure.

FIG. 3 illustrates a perspective view of a container, according to someembodiments of the present disclosure.

FIG. 4 illustrates a sectional view of the container of FIG. 3,according to some embodiments of the present disclosure.

FIG. 5 illustrates a side view of a dispenser, according to someembodiments of the present disclosure.

FIG. 6 illustrates a perspective view of the dispenser of FIG. 5 and thecontainer of FIG. 3, according to some embodiments of the presentdisclosure.

FIG. 7 illustrates a view of the sample device of FIG. 1 receiving asample, according to some embodiments of the present disclosure.

FIG. 8 illustrates a view of the sample device of FIG. 7, according tosome embodiments of the present disclosure.

FIG. 9 illustrates a perspective view of the sample device of FIG. 8 andthe container of FIG. 3, according to some embodiments of the presentdisclosure.

FIG. 10 illustrates a perspective view of the sample device and thecontainer of FIG. 9, according to some embodiments of the presentdisclosure.

FIGS. 11A-11B illustrate perspective views of the sample device and thecontainer, according to some embodiments of the present disclosure.

FIG. 12 illustrates a side view of a collection device, according tosome embodiments of the present disclosure.

FIG. 13 illustrates a side view of a sample device, according to someembodiments of the present disclosure.

FIG. 14 illustrates a sectional view of the sample device of FIG. 13,according to some embodiments of the present disclosure.

FIG. 15 illustrates a perspective view of a container, according to someembodiments of the present disclosure.

FIG. 16 illustrates a view of the collection device of FIG. 12 receivinga sample, according to some embodiments of the present disclosure.

FIG. 17 illustrates a view of the collection device of FIG. 16 and thesample device of FIGS. 13 and 14, according to some embodiments of thepresent disclosure.

FIG. 18 illustrates a view of the sample device of FIG. 17 and thecontainer of FIG. 15, according to some embodiments of the presentdisclosure.

FIG. 19 illustrates a view of the sample device and the container ofFIG. 18, according to some embodiments of the present disclosure.

FIG. 20 illustrates a view of the sample device and the container ofFIG. 19, according to some embodiments of the present disclosure.

FIGS. 21A-21B illustrate views of the sample device and the container ofFIG. 20, for dispensing of all or a portion of the contents, accordingto some embodiments of the present disclosure.

FIG. 22 illustrates a perspective view of a sample device, according tosome embodiments of the present disclosure.

FIG. 23 illustrates a sectional view of the sample device of FIG. 22,according to some embodiments of the present disclosure.

FIGS. 24-26 illustrate sectional views of the sample device of FIG. 22and a container, according to some embodiments of the presentdisclosure.

DETAILED DESCRIPTION

Various aspects now will be described more fully hereinafter. Suchaspects may, however, be embodied in many different forms and should notbe construed as limited to the embodiments set forth herein; rather,these embodiments are provided so that this disclosure will be thoroughand complete, and will fully convey its scope to those skilled in theart.

The cellular components of whole blood, and especially the red bloodcells, are the primary interfering substances in assays for a solubleconstituent of whole blood. Many blood tests are chromogenic, wherein asoluble constituent of the whole blood interacts with a particularreagent either to form a uniquely-colored complex or derivative as aqualitative indication of the presence or absence of the constituent, orto form a colored complex or derivative of variable color intensity as aquantitative indication of the presence of the constituent. The deep redcolor of the whole blood sample substantially interferes with thesechromogenic tests, and therefore the highly-colored red blood cellsusually are separated from the plasma or serum before the blood sampleis assayed for a particular soluble constituent. The presence of redblood cells also can interfere with various nonchromogenic blood assays,whereby the assay results are either inconsistent or, if consistent, areinaccurate. Furthermore, other cellular components, including the whiteblood cells, also can interfere in standard chromogenic blood assays.

Conventionally, plasma or serum can be separated from the cellularmaterial of whole blood by centrifugation or by clotting. The cellularmaterial collects at the bottom of the centrifuge or sample tube and thesupernatant plasma or serum is decanted. Accordingly, the interferingcellular components of whole blood are sufficiently removed such that asubstantial background interference is avoided. However, the centrifugemethod requires a large blood sample, a long centrifuge time, andappropriate equipment. Furthermore, the centrifuge method requiresseveral manipulative steps. The clotting method of obtaining serumrequires a long time (30-60 minutes) for clotting to occur andcentrifugation may be required after clotting occurs.

Other techniques can involve test strips for lateral flow of samples foranalysis. However, in some designs, red blood cells form aggregates orblockages within the chemical strip that disrupt the lateral flow. Suchblockages often produce inaccurate and/or invalid results.

Improved techniques to efficiently separate and accurately assay smallvolumes of whole blood are described herein. A separation technique canshield the technician from contact with the blood sample, avoid timedelays, and yield accurate and reproducible results. Techniques based onthe present disclosure can allow the safe, accurate, and economicalassay of a whole blood, or other biological fluid, sample for aparticular soluble component to achieve essentially total separation ofthe cellular components of whole blood from the plasma or serum. It willbe appreciated that the systems and devices and methods described hereinare contemplated for use with a variety of fluid samples, and whilst thedescription herein uses blood as a model sample, the systems and devicesand methods may also be used with other fluid samples.

Embodiments of the present disclosure can include a single use,disposable device. In one embodiment, the device is a whole bloodseparator in a fully contained, squeezable container including adropper. Embodiments described herein for use with a blood sample caninclude (1) a capillary tube that accurately draws a target bloodvolume, (2) a pad that chemically interacts with red-blood cells, suchthat the red blood cells become chemically and/or physically trappedwithin the pad material, (3) a mechanism for plasma recovery from thepad upon diffusion or active mixing, and (4) a dropper tip thatfacilitates dispensing the mixture onto a test device. Embodiments ofthe present disclosure can incorporate a blood capillary tube with avolume monitoring mechanism. Embodiments of the present disclosure canfacilitate removal of red-blood cells from a patient venipuncture offinger stick whole blood sample, such that plasma and buffer solutioncan be added to the test strip. The design can simplify the work-flowfor the end-user and can be applied universally across assays thatutilize a whole blood sample requiring a plasma matrix.

Chemically or physically capturing the red-blood cells in the padmaterial improves the efficiency of plasma separation, specifically theefficiency of red blood cell capture in a pad material which requiresless solution volume to recover plasma (reduce sample dilution effect)and prevents red blood cells from lysing and/or leaking from the padonto the test device upon use. The treatment of the red blood cells canbe performed prior to contact with a buffer solution, so the red bloodcells will not be, or will be minimally, released into the buffersolution when the sample and buffer solution are mixed or when pressureis added to the container to express the sample solution.

A system can facilitate processing of a fluid or liquid sample, and inparticular a whole blood sample from a human patient or other mammal. Inone embodiment, the system is for use with a whole blood sample and, aswill be shown, it effects separation and dispensation of the plasmawhile the cellular components are retained in the system. The system caninclude a sample device for receiving and treating a liquid sample froma patient. FIGS. 1 and 2 illustrate a sample device 100, according tosome embodiments of the present disclosure. According to someembodiments, for example as illustrated in FIGS. 1 and 2, the sampledevice 100 can include an inflow chamber 110 for receiving a sample. Theinflow chamber 110 can include a capillary tube with an open end locatedon a first side of the sample device 100. The inflow chamber 110 can beseparated from a mixing chamber 130 by a pad 140. The pad 140 canoptionally include a processing reagent, such as an anti-RBC reagent,such as an anti-human red blood cell antibody that is configured tofacilitate collection of blood cells therein. The pad 140 can be a fiberor polymer membrane which has pore size larger than a cellularcomponent, e.g., red blood cells, so that the red blood cells enterfreely, but smaller than a processed cellular component, e.g.,agglutinated red blood cells (e.g., agglutinated by the interaction withanti-human RBC antibody). The sample device 100 can also include achannel 150 that provides fluid communication to the mixing chamber 130via a pathway other than through the inflow chamber 110 and the pad 140.As described further herein, the channel 150 provides a pathway forreagents after connecting the sample device 100 to a container holdingthe reagents.

The sample device 100 can also include a filter 160 between the mixingchamber 130 and an outflow port 190. The filter 160 can be configured toallow passage of a first fraction or portion of the liquid sample, suchas a plasma fraction of a blood sample, there through while retaining asecond fraction or portion of the liquid sample, such as cellularcomponents of a whole blood sample. The filter 160 can include aseparation membrane (for example, polysulfone material) withasymmetrical porosity to trap a component of the sample. Alternativelyor in combination, the filter 160 can include a filter portion (polymer,glass fiber, woven mesh, etc.) with pore size smaller than a treated ornon-treated portion of the liquid sample, such as agglutinated red bloodcells, in order to trap any of this portion that may escape pad 140. Thethickness of the filter 160 may be selected to obtain the amount offiltration required while retaining a minimum volume of plasma orreagent. The outflow port 190 can be located at an end of the sampledevice 100 that is opposite the inflow chamber 110 and allowdispensation of a sample portion (e.g., a plasma portion of a bloodsample) that is passed from the filter 160. In some embodiments, thefilter 160 is not included, and a portion of the sample, (e.g., thecellular components) are retained within the pad 140. The sample device100 can include a device interface 120 (e.g., threads) that canfacilitate connection and securement of the sample device 100 to acontainer, as further described herein.

The system can include a container that helps facilitate mixing of asample with a reagent. FIGS. 3 and 4 illustrate a container 200,according to some embodiments of the present disclosure. According tosome embodiments, for example as illustrated in FIGS. 3 and 4, thecontainer 200 can include a reservoir 220 for holding and/or receiving areagent, such as a solution containing a liquid, for example aprocessing solution, water, saline, a buffer, phosphate buffered saline,an electrolytic solution, etc. The reservoir 220 can be placed in fluidcommunication with at least a portion of the sample device 100, so thata fluid (liquid) from the sample device 100 and a fluid (liquid) fromthe container 200 can mix. For example, the reservoir 220 can be placedin fluid communication with the mixing chamber 130 of the sample device100. The container 200 can include a container interface 210 (e.g.,threads) that can facilitate connection and securement of the container200 to the sample device 100. For example, the sample device 100 can beconnected to the container 200 by engaging the device interface 120 withthe container interface 210. While engaged, the device interface 120 andthe container interface 210 maintain a connection between the sampledevice 100 and the container 200. The device interface 120 and thecontainer interface 210 can also seal a space within the sample device100 and the container 200 so that materials within the space areseparate from an external environment. At least a portion of thecontainer 200 can be deformable, to allow a user to squeeze thecontainer 200 and dispense a fluid, as described further herein.

The system can include substances for use with other components of thesystem to facilitate separation of the liquid sample into a firstportion and a second portion, such as required in whole bloodseparation. FIG. 5 illustrates a reagent 20, such as a solutioncontaining a solution, within a dispenser 300, according to someembodiments of the present disclosure. According to some embodiments,for example as illustrated in FIG. 5, the dispenser 300 can include areservoir 310 for holding and/or dispensing a fluid. For example, thedispenser 300 can be ampoule with a predetermined amount of a reagent.The reservoir 310 can be initially sealed by a removable cap 330.

According to embodiments, a kit of parts can include components of thesystem. For example, one or more kits of parts can include the sampledevice 100, the container 200, and/or the dispenser 300. Directions foruse (“DFU”) can be included with the kit. One or more of the sampledevice 100, the container 200, and the dispenser 300 can be disposableand packaged for one-time use.

A method can employ components of the system or kit, as describedherein, to achieve processing of a liquid sample, such as whole bloodseparation. According to some embodiments, for example as illustrated inFIG. 6, the reagent 20 can be dispensed from the dispenser 300 and intothe reservoir 220 of the container 200. Alternatively, the container 200is configured to contain a liquid, and the reservoir of the containercan be filled with a liquid by an end user of the system or kit.

According to some embodiments, for example as illustrated in FIG. 7, asample 10, such as blood, can be collected with and into the inflowchamber 110 of the sample device 100. The sample device 100 can be heldhorizontally or substantially horizontally during filling of the inflowchamber 110. The inflow chamber 110 can automatically cease to collectthe sample 10 once the inflow chamber 110 is filled. For example, whenthe inflow chamber is configured to be a capillary tube, collection canstop when the capillary is full, such that there is no surface left forthe liquid sample (e.g., blood) to flow into. At such a saturationpoint, the inflow chamber 110 will not absorb any more liquid.Accordingly, in one embodiment, the inflow chamber of the sample deviceis configured as a capillary tube with a defined length that collects afixed volume of a fluid sample.

According to some embodiments, for example as illustrated in FIG. 8, thesample device 100 can be positioned vertically with the inflow chamber110 gravitationally above the mixing chamber 130 (not visible in FIG.8). While the sample device 100 is in such a position, the sample 10 canbe allowed to drain from the inflow chamber 110 and to the pad 140and/or the mixing chamber 130 (e.g., via the pad 140). As the sample 10passes through the pad 140, the sample 10 can react with substanceswithin the pad 140, such as anti-RBC antibodies. For example, at least aportion of the sample 10 (e.g., plasma or serum) can be separated from asecond portion (e.g., red blood cells) of the sample 10 by retaining thesecond portion (e.g., red blood cells) at the pad 140. The sample device100 can be held vertically for an amount of time sufficient to allow thesample 10 to drain out of this inflow chamber 110 (e.g., 10 seconds, 20seconds, 30 seconds, 40 seconds, 50 seconds, 60 seconds, or more than 60seconds).

According to some embodiments, for example as illustrated in FIG. 9, thesample device 100 can be connected to the container 200. For example,the inflow chamber 110 can be inserted into the reservoir 220 of thecontainer 200. The device interface 120 can engage the containerinterface 210 to seal a space between the sample device 100 and thecontainer 200. The sample device 100 can be connected to the container200 in a way that allows the outflow port 190 to be exposed orexposable.

According to some embodiments, for example as illustrated in FIG. 10,the sample device 100 and the container 200 can be agitated and/orshaken to mix at least a portion of the sample 10 in the sample device100 with at least a portion of the reagent 20 in the container 200. Themixing can occur, at least in part, within the mixing chamber 130. Forexample, during the mixing, the reagents 20 can pass through the channel150 to the mixing chamber 130. Within the mixing chamber 130, thereagents can mix with the sample 10 that is on or within the pad 140.The sample device 100 and the container 200 can be agitated and/orshaken manually or by a machine.

According to some embodiments, for example as illustrated in FIGS.11A-11B, the mixture produced by the agitation and/or shaking can bedispensed from the sample device 100 and the container 200. The outflowport 190 can be directed toward a receptacle 410 of a cassette 400. Inone embodiment, at least a portion of the container 200 and/or thesample device 100 can be deformed to create pressure within the mixingchamber 130, as indicated in FIG. 11A by the arrows. In response to thisincreased pressure, a portion of the mixture 30 within the mixingchamber 130 can be forced through the filter 160 and dispensed from theoutflow port 190. At least a portion of the mixture 30 can be retainedwithin the mixing chamber 130. For example, the cellular components thatare too large to fit through the filter 160 can be retained within themixing chamber 130 while the plasma passes through the filter 160 andthe outflow port 190. In another embodiment, as illustrated in FIG. 11B,sample device 100 comprises a skirt 193 surrounding outflow port 190,the skirt configured to engage with an external docking station orsample receiving zone, such as receptacle 410. Mixture 30 is dispensedfrom the mixing chamber by applying a directional force to the sampledevice or the container to engage or actuate a mechanism to release themixture. In one embodiment, the directional force is, with respect to agravitational frame of reference, a downward force. The downward appliedforce effects release of the fluid mixture from the sample device.Mechanisms responsive to a downward force applied by a user are referredto as “press and release” or “press to dispense” mechanisms. In oneembodiment, dispensing mixture from the mixing chamber comprisesapplying a directional force to effect release of mixture, whereinsubstantially all of the mixture in the mixing chamber is dispensed. Inother embodiments, greater than 90%, 95%, 98% or 99% of the mixture inthe mixing chamber is dispensed.

In one embodiment, a portion or all of an inner surface of outflow port190 is treated with material that facilitates release of mixture. Forexample, a hydrophilic material or a hydrophobic material can be appliedto or coated on the inner surface of the outflow port, the container,the mixing chamber, or other surfaces of the sample device andcontainer, to modify the surface-liquid mixture surface tension.

Alternatively or in combination with the above embodiments, systems canfacilitate processing of a fluid sample, such as a whole blood sample orsputum sample or urine sample, to separate the sample into a firstportion, such as plasma or protein free sputum or urine, and a secondsample portion, such as cellular components of blood, protein or cellsof sputum or urine, and dispensing of the first portion only from thesystem—that is, the second portion is retained in the system. Accordingto some embodiments, such a system can include a sample device forreceiving a sample from a patient. FIG. 12 illustrates a collectiondevice 500, according to some embodiments of the present disclosure.According to some embodiments, for example as illustrated in FIG. 12,collection device 500 can include an inflow chamber 510 for receiving asample. The inflow chamber 510 can include a capillary tube with an openend located on a first side of the collection device 500, however itwill be appreciated that the inflow chamber need to be a capillary tubebut may be of other configurations and geometries. The inflow chamber510 can be in fluid communication with a compression chamber 520. Atleast a portion of collection device 500 can be deformable, tofacilitate collection of the sample within the inflow chamber undervarious pressure conditions. The collection device 500 can include amarker 530 to provide a target fill indication along the length ofinflow chamber 510. In one embodiment, where the inflow chamber isconfigured as a capillary tube, the inflow chamber has a defined lengthfor collection of a fixed, known, and/or predetermined volume of sample.

The system can include a sample device for receiving a sample from apatient and/or receiving a sample from the collection device. FIG. 13illustrates a sample device 600, according to some embodiments of thepresent disclosure. According to some embodiments, for example asillustrated in FIGS. 13 and 14, the sample device 600 can include a pad610 for receiving a sample. The pad 610 can include a processingreagent, such as an anti-RBC reagent that is configured to facilitatecollection of blood cells therein. The sample device 600 can alsoinclude a chamber 630 (visible in FIG. 14) for receiving the sampleand/or reagents when connected to a container, as described furtherherein.

With reference to FIG. 14, the sample device 600 can also include afilter 660 between chamber 630 and an outflow port 690. Filter 660 canbe configured to allow passage of a first fraction of the sample, suchas plasma, there through while retaining a second fraction of thesample, such as cellular components of the sample. In one embodiment,the filter is a glass fiber filter. Outflow port 690 can be located atan end of sample device 600 that is opposite pad 610 and allowdispensation of that fraction or portion of sample, such as the plasma,that is passed from filter 660. In some embodiments, filter 660 is notincluded, and the cellular components are retained within pad 610. Thesample device 600 can include a device interface 620 (e.g., threads)that can facilitate connection and securement of sample device 600 to acontainer, as further described herein.

The system can include a container that helps facilitate mixing of asample with a reagent. FIG. 15 illustrates a container 700, according tosome embodiments of the present disclosure. According to someembodiments, for example as illustrated in FIG. 15, container 700 caninclude a reservoir 720 for holding and/or receiving a fluid or areagent, such as a solution containing a buffer. Reservoir 720 can beplaced in fluid communication with at least a portion of sample device600, so that a fluid from sample device 600 and a fluid from container700 can mix. For example, reservoir 720 can be placed in fluidcommunication with chamber 630 of the sample device 600. The container700 can include a container interface 710 (e.g., threads) that canfacilitate connection and securement of container 700 to sample device600. For example, sample device 600 can be connected to container 700 byengaging device interface 620 with container interface 710. Whileengaged, device interface 620 and container interface 710 maintain aconnection, preferably a fluid-tight connection, between the sampledevice and the container. The device interface 620 and the containerinterface 710 can also seal a space within the sample device 600 and thecontainer 700 so that materials within the space are separate from anexternal environment. In one embodiment, at least a portion of thecontainer 700 can be deformable, to allow a user to squeeze thecontainer 700 and dispense a fluid, as described further herein.

According to embodiments, a kit of parts can include components of thesystem. For example, one or more kits of parts can include thecollection device 500, the sample device 600, and/or the container 700.Directions for use (“DFU”) can be included with the kit. One or more ofthe collection device 500, the sample device 600, and the container 700can be disposable and packaged for one-time use.

A method can employ components of the system or kit, as describedherein, to achieve processing of a fluid sample, such as processing of awhole blood sample into two portions. According to some embodiments, forexample as illustrated in FIG. 16, a sample 10, such as blood, can becollected with and into the inflow chamber 510 of the collection device500. The collection device 500 can be held horizontally or substantiallyhorizontally during filling of the inflow chamber 510. The compressionchamber 520 can be compressed prior to collection and then released toreduce the pressure at the inflow chamber 510 and create flow into theinflow chamber 510. Collection can continue until the sample 10 reachesthe marker 530.

According to some embodiments, for example as illustrated in FIG. 17,the collection device 500 can be positioned to dispense all or a portionof the sample 10 to the pad 610 of the sample device 600. Thecompression chamber 520 can be compressed to increase the pressure atthe inflow chamber 510 and create flow out of the inflow chamber 510.The sample 10 can be absorbed into the pad 610. In one embodiment, thecollection device is configured to collect a fixed, known, and/ordetermined volume of sample. In one embodiment, greater than 90%, 95%,98% or 99% of the sample in the inflow chamber of the collection deviceis dispensed onto pad 610.

According to some embodiments, for example as illustrated in FIGS. 18and 19, the sample device 600 can be connected to the container 700. Forexample, the pad 610 can be inserted into the reservoir 720 of thecontainer 700. The device interface 620 can engage the containerinterface 710 to seal a space between the sample device 600 and thecontainer 700. The sample device 600 can be connected to the container700 in a way that allows the outflow port 690 to be exposed orexposable.

According to some embodiments, for example as illustrated in FIG. 20,the sample device 600 and the container 700 can be agitated and/orshaken to mix at least a portion of the sample 10 in the sample device600 with at least a portion of the reagents 20 (not shown in FIGS. 18and 19) in the container 700. The mixing can occur, at least in part,within the chamber 630. Within the chamber 630 and/or the reservoir 720,the reagents 20 can mix with the sample 10 that is on or within the pad610. The sample device 600 and the container 700 can be agitated and/orshaken manually or by a machine.

According to some embodiments, for example as illustrated in FIGS.21A-21B, the mixture produced by the agitation and/or shaking can bedispensed from the sample device 600 and the container 700. The outflowport 690 can be directed toward a receptacle 810 of a cassette 800. Inone embodiment, as shown in FIG. 21A, at least a portion of thecontainer 700 and/or the sample device 600 can be deformed to createpressure within the chamber 630. In response to this increased pressure,a portion of the mixture 30 within the chamber 630 can be forced throughthe filter 660 and dispensed from the outflow port 690. At least aportion of the mixture 30 can be retained within the chamber 630. Forexample, the cellular components that are too large to fit through thefilter 660 can be retained within the chamber 630, while the plasmapasses through the filter 660 and the outflow port 690. In anotherembodiment, shown in FIG. 21B, sample device 600 comprises a skirt 693surrounding outflow port 690, the skirt configured to engage with anexternal docking station or sample receiving zone, such as receptacle810. The fluid mixture in the chamber of the sample device is dispensedby applying a directional force to the sample device, or to the attachedcontainer, to engage or actuate a mechanism to release the mixture. Inone embodiment, the directional force is, with respect to agravitational frame of reference, a downward force. The downward appliedforce effects release of the fluid mixture from the sample device.Mechanisms responsive to a downward force applied by a user are referredto as “press and release” or “press to dispense” mechanisms. In oneembodiment, dispensing mixture from chamber 630 comprises applying adirectional force to effect release of mixture, wherein substantiallyall of the mixture in the chamber is dispensed. In other embodiments,greater than 90%, 95%, 98% or 99% of the mixture in the chamber isdispensed.

A system can facilitate whole blood separation and dispensation of theplasma while the cellular components are retained. The system caninclude a sample device for receiving and treating a sample from apatient. FIGS. 22 and 23 illustrate a sample device 950, according tosome embodiments of the present disclosure. According to someembodiments, for example as illustrated in FIGS. 22 and 23, the sampledevice 950 can include an inflow chamber 960 for receiving a sample. Theinflow chamber 960 can include a capillary tube with an open end locatedon a first side of the sample device 950. The inflow chamber 960 can beseparated from and/or divide channels 990 with a pad 980. The pad 980can include an anti-RBC reagent that is configured to facilitatecollection of blood cells therein. The channels 990 can provide accessto the pad 980 on one or more sides of the pad 980. As described furtherherein, the channels 990 provide a pathway for reagents after activatingthe sample device 950 within a container holding the reagents.

The system can include a container that helps facilitate mixing of asample with a reagent. FIGS. 24-26 illustrate a container 900, accordingto some embodiments of the present disclosure. According to someembodiments, for example as illustrated in FIG. 24, the container 900can include a reservoir 940 for holding and/or receiving a reagent 20,such as a solution containing a buffer. The reservoir 940 can be sealed,at least in part, with a barrier 930, such as a foil cover. The barrier930 can isolate the reservoir 940 from a chamber 920 of the container900 until the barrier 930 is punctured, broken, or removed.

The sample device 950 can be placed at least partially within thecontainer 900. For example, the container 900 can include protrusions922 that can secure at least a portion of the sample device 950. Forexample, one or more protrusions 922 can be placed on opposing sides ofa portion of the sample device 950, such that the sample device 950 issecured within the container 900 until an external force moves thesample device 950 past one or more of the protrusions 922.

A method can employ components of the system or kit, as describedherein, to achieve whole blood separation. According to some embodimentsa sample, such as blood, can be collected with and into the inflowchamber 960 of the sample device 950. The sample device 950, whilesecured within the container 900, can be held horizontally orsubstantially horizontally during filling of the inflow chamber 960.

According to some embodiments, for example as illustrated in FIG. 24,the sample device 950 can be positioned vertically with the inflowchamber 960 gravitationally above the pad 980. While the sample device950 is in such a position, the sample can be allowed to drain from theinflow chamber 960 and to the pad 980 and/or the channels 990 (e.g., viathe pad 980). As the sample passes through the pad 980, the sample canreact with substances within the pad 980, such as anti-RBC reagents. Forexample, at least a portion of the sample can be separated from a secondportion of the sample by retaining the second portion at a pad 980. Thesample device 950 can be held vertically for an amount of timesufficient to allow the sample to drain out of this inflow chamber 960(e.g., 10 seconds, 20 seconds, 30 seconds, 40 seconds, 50 seconds, 60seconds, or more than 60 seconds).

According to some embodiments, for example as illustrated in FIGS.24-26, the sample device 950 can be advanced within the container 900 torelease a reagent. For example, an end 992 of the sample device 950 canpuncture the barrier 930 and be at least partially inserted into thereservoir 940 of the container 900. The sample device 950 can beadvanced by overcoming a force provided by the one or more protrusions922 that support the sample device 950. For example, a dispenser 912 canbe used to advance the sample device 950. A dispenser interface 918 canengage a container interface 910 to seal a space (e.g., the chamber 920)within the container 900. For example, the container 900 can include thecontainer interface 910 (e.g., threads) that can facilitate connectionand securement of the container 900 to the sample device 950. The sampledevice 950 can be connected to the container 900 by engaging thedispenser interface 918 with the container interface 910. While engaged,the dispenser interface 918 and the container interface 910 maintain aconnection between the dispenser 912 and the container 900. Thedispenser interface 918 and the container interface 910 can also seal aspace within the container 900 so that materials within the space areseparate from an external environment. As the dispenser 912 is attached,it can apply a force to the sample device 950 to urge the end 992 of thesample device 950 through the barrier 930. In such a configuration, thereagent 20 is allowed to mix with the sample within the pad 980. Forexample, the reagent 20 can travel within the channels 990 to access thepad 980 and/or the chamber 920.

According to some embodiments, the sample device 950 and the container900 can be agitated and/or shaken to mix at least a portion of thesample in the sample device 950 with at least a portion of the reagent20 in the container 900. The mixing can occur, at least in part, withinthe channels 990. Within the channels 990, the reagents can mix with thesample that is on or within the pad 980. The sample device 950 and thecontainer 900 can be agitated and/or shaken manually or by a machine.

According to some embodiments, the mixture produced by the agitationand/or shaking can be dispensed from the sample device 950 and thecontainer 900 via the dispenser 912. An outflow port 916 of thedispenser 912 can be directed toward a receptacle of a cassette (notshown). At least a portion of the container 900 can be deformed tocreate pressure within the channels 990. In response to this increasedpressure, a portion of the mixture within the container 900 can beforced through a filter 914 of the dispenser 912 and dispensed from theoutflow port 916. The filter 914 can include, for example, glass fibers,and be configured to allow passage of plasma there through whileretaining cellular components of the sample. At least a portion of themixture can be retained within the channels 990. For example, thecellular components that are too large to fit through the filter 914 canbe retained within the channels 990 while the plasma passes through thefilter 914 and the outflow port 916. In some embodiments, the filter 914is not included, and the cellular components are retained within the pad980.

According to embodiments, a kit of parts can include components of thesystem. For example, one or more kits of parts can include the sampledevice 950, the container 900, and the dispenser 912. Directions for use(“DFU”) can be included with the kit. One or more of the sample device950, the container 900, and the dispenser 912 can be disposable andpackaged for one-time use.

Examples

A study was performed to evaluate the use of the described whole bloodseparation device with finger stick whole blood compared to the serumand plasma testing methods using the FDA-cleared SOFIA® Lyme FIA testdevice from Quidel Corporation. This prospective study was performedusing matched blood specimens (finger stick and serum/plasma) samplescollected from 529 subjects suspected of and exhibiting symptoms of Lymedisease across 11 sites located in Lyme endemic regions throughout theUnited States. The whole blood sampling and Lyme immunoassay wereperformed immediately at each of the 11 clinical sites by CLIA-waivedoperators, which demonstrates the ease of use in the hands ofnon-trained operators. The predicate Lyme IgM and IgG assays and westernblot Lyme IgM and IgG assays were performed at central referencelaboratories that were different from the CLIA-waived testing sites.First tier results of the whole blood separation device with fingerstick whole blood compared to no separation device plasma and serumusing the SOFIA® Lyme IgM and IgG assay are shown in Tables 1 and 2.There is no statistical difference (p>0.05) between conditions,indicating that the whole blood separation device performs as well asthe tests run with plasma and serum.

TABLE 1 Comparison of a whole blood separation device (finger stickwhole blood) to no separation device (plasma and serum) using theSOFIA ® Lyme IgM FIA prospective study positive and negative results.IgM Results (Sofia) Separation Device (Finger stick No Sep Device No SepDevice blood) (Plasma) (Serum) Positive 238 243 241 Negative 291 286 288

TABLE 2 Comparison of a whole blood separation device (finger stickwhole blood) to no separation device (plasma and serum) using the SofiaLyme IgG FIA prospective study positive and negative results. IgGResults (SOFIA ®) Separation Device (Finger stick No Sep Device No SepDevice blood) (Plasma) (Serum) Positive 203 198 192 Negative 326 331 337

Clinical performance was evaluated based on a comparison of the SofiaLyme FIA results when testing capillary whole blood to those obtained bythree different Tier 1 IgM and IgG comparator methods testing a matchedserum or plasma specimen. The SOFIA® Lyme NM and IgG percent positiveand percent negative agreement to the comparator test devices wereevaluated using the whole blood separation device with finger stickwhole blood compared to no separation device plasma and serum, This datais presented in Tables 3 and 4. There is no statistical difference(p>0.05) between conditions, indicating that the whole blood separationdevice performs as well as the tests run with plasma and serum.

TABLE 3 Comparison of whole blood separation device (finger stick wholeblood) to no separation device (plasma and serum) using the SOFIA ® LymeIgM FIA prospective study percent positive and percent negativeagreement to the first Tier comparator methods. IgM Agreement toComparators (SOFIA ®) Separation Device (Finger No Sep Device No SepDevice stick blood) (Plasma) (Serum) % Positive Agreement 79% 79% 79% %Negative 73% 71% 72% Agreement

TABLE 4 Comparison of whole blood separation device (finger stick wholeblood) to no separation device (plasma and serum) using the SOFIA ® LymeIgG FIA prospective study percent positive and percent negativeagreement to the first Tier comparator methods. IgG Agreement toComparators (SOFIA ®) Separation Device (Finger No Sep Device No SepDevice stick blood) (Plasma) (Serum) % Positive Agreement 90% 89% 89% %Negative 81% 82% 84% Agreement

Follow-up second tier testing with IgM and IgG Western blots wasperformed on all first tier positive results. Second Tier results wereevaluated using the whole blood separation device with finger stickwhole blood compared to no separation device plasma and serum. This datais presented in Table 5. There is no statistical difference (p>0.05)between conditions for both IgM and IgG second tier positive results,indicating that the whole blood separation device performs as well asthe tests run with plasma and serum.

TABLE 5 Comparison of a whole blood separation device (finger stickwhole blood) to no separation device (plasma and serum) using theSOFIA ® Lyme IgM and IgG FIA prospective study results that areconfirmed positive by Western Blot. Second Tier Western BlotConfirmation Separation Device (Finger No Sep Device No Sep Device stickblood) (Plasma) (Serum) IgM Positive Results 106 105 106 IgG PositiveResults 72 73 71

Follow-up second tier testing with IgM and IgG Western blots wasperformed on first tier positive results generated with the SOFIA® wholeblood separation device and all comparator methods that did not use thewhole blood separation device. For Lyme IgM, the results demonstratethat there are more second tier positive results using the SOFIA® wholeblood separation device (106) compared to the comparator methods(positive results range between 77-95). This means that there are moresuspected Lyme positive patients in the early stage of disease that willbe diagnosed using the SOFIA® IgM whole blood method. For Lyme IgG, theresults demonstrate that there are a similar number of second tierpositive results using the Sofia whole blood separation device (72)compared to the comparator methods (positive results range between57-72). This means that in some cases there are more suspected Lymepositive patients that will be diagnosed using the SOFIA® IgG wholeblood method. The results are summarized in Table 6.

TABLE 6 Comparison of a whole blood separation device (finger stickwhole blood) to no separation device (plasma and serum) using theSOFIA ® Lyme IgM and IgG FIA prospective study results that areconfirmed positive by Western Blot. Second Tier Western BlotConfirmation Test Method IgM Positive Results IgG Positive Results SofiaSep Device 106 72 (Finger stick blood) Biorad (Serum) 82 72 Trinity(Serum) 80 57 Zeus (Serum) 95 72 Vidas (Serum) 77 70

The described whole blood separation device has a number of advantagesover the traditional serum and plasma method. For example, the describedwhole blood separation allows a small sample volume (25 ul blood from afinger stick instead of large volume drawn from venipuncture tube). Byfurther example, the described whole blood separation allows quickturn-around time (<60 seconds) compared to centrifugation (>15 min). Byfurther example, the described whole blood separation allows ease of usecompared to centrifugation, thereby allowing use by CLIA waivedoperators instead of trained lab operators. By further example, thedescribed whole blood separation requires no additional equipment,providing a single-use separation device instead of lab instruments,such as a centrifuge. By further example, the described whole bloodseparation provides similar first tier performance compared to serum andplasma Lyme IgM and IgG results. By further example, the described wholeblood separation provides improved second tier performance compared totraditional methods for Lyme IgM and IgG.

The foregoing description is provided to enable a person skilled in theart to practice the various configurations described herein. While thesubject technology has been particularly described with reference to thevarious figures and configurations, it should be understood that theseare for illustration purposes only and should not be taken as limitingthe scope of the subject technology.

A phrase such as “an aspect” does not imply that such aspect isessential to the subject technology or that such aspect applies to allconfigurations of the subject technology. A disclosure relating to anaspect may apply to all configurations, or one or more configurations.An aspect may provide one or more examples of the disclosure. A phrasesuch as “an aspect” may refer to one or more aspects and vice versa. Aphrase such as “an embodiment” does not imply that such embodiment isessential to the subject technology or that such embodiment applies toall configurations of the subject technology. A disclosure relating toan embodiment may apply to all embodiments, or one or more embodiments.An embodiment may provide one or more examples of the disclosure. Aphrase such “an embodiment” may refer to one or more embodiments andvice versa. A phrase such as “a configuration” does not imply that suchconfiguration is essential to the subject technology or that suchconfiguration applies to all configurations of the subject technology. Adisclosure relating to a configuration may apply to all configurations,or one or more configurations. A configuration may provide one or moreexamples of the disclosure. A phrase such as “a configuration” may referto one or more configurations and vice versa.

There may be many other ways to implement the subject technology.Various functions and elements described herein may be partitioneddifferently from those shown without departing from the scope of thesubject technology. Various modifications to these configurations willbe readily apparent to those skilled in the art, and generic principlesdefined herein may be applied to other configurations. Thus, manychanges and modifications may be made to the subject technology, by onehaving ordinary skill in the art, without departing from the scope ofthe subject technology.

It is understood that the specific order or hierarchy of steps in theprocesses disclosed is an illustration of exemplary approaches. Basedupon design preferences, it is understood that the specific order orhierarchy of steps in the processes may be rearranged. Some of the stepsmay be performed simultaneously. The accompanying method claims presentelements of the various steps in a sample order, and are not meant to belimited to the specific order or hierarchy presented.

As used herein, the phrase “at least one of” preceding a series ofitems, with the term “and” or “or” to separate any of the items,modifies the list as a whole, rather than each member of the list (i.e.,each item). The phrase “at least one of” does not require selection ofat least one of each item listed; rather, the phrase allows a meaningthat includes at least one of any one of the items, and/or at least oneof any combination of the items, and/or at least one of each of theitems. By way of example, the phrases “at least one of A, B, and C” or“at least one of A, B, or C” each refer to only A, only B, or only C;any combination of A, B, and C; and/or at least one of each of A, B, andC.

Terms such as “top,” “bottom,” “front,” “rear” and the like as used inthis disclosure should be understood as referring to an arbitrary frameof reference, rather than to the ordinary gravitational frame ofreference. Thus, a top surface, a bottom surface, a front surface, and arear surface may extend upwardly, downwardly, diagonally, orhorizontally in a gravitational frame of reference.

Furthermore, to the extent that the term “include,” “have,” or the likeis used in the description or the claims, such term is intended to beinclusive in a manner similar to the term “comprise” as “comprise” isinterpreted when employed as a transitional word in a claim.

The word “exemplary” is used herein to mean “serving as an example,instance, or illustration.” Any embodiment described herein as“exemplary” is not necessarily to be construed as preferred oradvantageous over other embodiments.

A reference to an element in the singular is not intended to mean “oneand only one” unless specifically stated, but rather “one or more.”Pronouns in the masculine (e.g., his) include the feminine and neutergender (e.g., her and its) and vice versa. The term “some” refers to oneor more. Underlined and/or italicized headings and subheadings are usedfor convenience only, do not limit the subject technology, and are notreferred to in connection with the interpretation of the description ofthe subject technology. All structural and functional equivalents to theelements of the various configurations described throughout thisdisclosure that are known or later come to be known to those of ordinaryskill in the art are expressly incorporated herein by reference andintended to be encompassed by the subject technology. Moreover, nothingdisclosed herein is intended to be dedicated to the public regardless ofwhether such disclosure is explicitly recited in the above description.

While certain aspects and embodiments of the subject technology havebeen described, these have been presented by way of example only, andare not intended to limit the scope of the subject technology. Indeed,the novel methods and systems described herein may be embodied in avariety of other forms without departing from the spirit thereof. Theaccompanying claims and their equivalents are intended to cover suchforms or modifications as would fall within the scope and spirit of thesubject technology.

What is claimed is:
 1. A kit comprising: a container comprising areservoir; a sample device comprising: an inflow chamber for receiving afluid sample; a mixing chamber; a pad between the inflow chamber and themixing chamber, the pad comprising a processing reagent; an outflow portadjacent to the mixing chamber; and an interface for sealably connectingthe sample device to the container such that the reservoir is in fluidcommunication with the mixing chamber.
 2. The kit of claim 1, whereinthe inflow chamber comprises a capillary tube.
 3. The kit of claim 2,wherein the capillary tube has a length selected to collect a fixedvolume of fluid sample.
 4. The kit of claim 1, wherein the sample devicefurther comprises a filter between the mixing chamber and the outflowport.
 5. The kit of claim 1, further comprising a buffer solution withinthe reservoir.
 6. The kit of claim 1, wherein the processing reagent isa red blood cell agglomerating substance.
 7. A kit comprising: a sampledevice comprising: a pad for receiving a liquid sample; and a chamberhaving an outflow port; and a container comprising: a reservoirconfigured to contain or containing a liquid solution; and an interfacefor sealably connecting the sample device to the container such that thereservoir is in fluid communication with the chamber.
 8. The kit ofclaim 7, further comprising a collection device for collecting theliquid sample.
 9. The kit of claim 7, wherein the collection devicecomprises a capillary tube and a bulb in fluid communication with thecapillary tube.
 10. The kit of claim 7, wherein the pad comprises aprocessing reagent.
 11. The kit of claim 10, wherein the processingreagent is a red blood cell agglomerating substance.
 12. A kitcomprising: a container comprising: a reservoir containing or configuredto contain a solution; a mixing chamber; and a barrier separating thereservoir from the mixing chamber; a sample device comprising: an inflowchamber for receiving a liquid sample; a channel; and a pad between theinflow chamber and the channel; and a dispenser comprising: an outflowport; and an interface for sealably connecting the dispenser to thecontainer such that the mixing chamber is in fluid communication withthe outflow port, wherein the dispenser is configured to urge the sampledevice through the barrier when the dispenser is connected to thecontainer.
 13. The kit of claim 12, wherein the inflow chamber comprisesa capillary tube.
 14. The kit of claim 12, wherein the dispenser furthercomprises a filter that is between the mixing chamber and the outflowport when the dispenser is connected to the container.
 15. The kit ofclaim 14, wherein the filter is positioned between opposing openings ofthe dispenser.
 16. The kit of claim 12, wherein the container comprisesat least one protrusion to secure at least a portion of the sampledevice within the mixing chamber.
 17. The kit of claim 16, wherein thedispenser is configured to urge the at least a portion of the sampledevice through the at least one protrusion when the dispenser isconnected to the container.
 18. The kit of claim 12, wherein thecontainer comprises multiple protrusions to secure at least a portion ofthe sample device between the multiple protrusions and within the mixingchamber.
 19. The kit of claim 12, wherein the pad comprising aprocessing reagent.
 20. The kit of claim 19, wherein the processingreagent is a red blood cell agglomerating substance.